Use of bioactive fraction from cow urine distillate (‘Go-mutra’) as a bio-enhancer of anti-infective, anti-cancer agents and nutrients

ABSTRACT

The invention relates to a novel pharmaceutical composition comprising an effective amount of bio-active fraction from cow urine distillate as a bioavailability facilitator and pharmaceutically acceptable additives selected from anticancer compounds, antibiotics, drugs, therapeutic and nutraceutic agents, ions and similar molecules which are targeted to the living systems.

This application is a divisional, of application Ser. No. 10/135,763,filed May 1, 2002, now U.S. Pat. No. 6,896,907 which is a divisional ofapplication Ser. No. 09/726,307, filed Dec. 1, 2000, now U.S. Pat. No.6,410,059. issued Jun. 25, 2002, which are both hereby expresslyincorporated by reference in their entirety.

This application claims priority under 35 U.S.C. §§ 119 and/or 365 to60/241,842 filed in the U.S. on Oct. 20, 2000; the entire content ofwhich is hereby incorporated by reference

FIELD OF THE INVENTION

The invention relates to an absolutely novel use of cow urine distillateas activity enhancer and availability facilitator for bioactivemolecules including anti-infective and anti-cancer agents. The moleculeswhich express any activity in form of either inhibiting or promoting abiological function have been referred in this invention as bioactivemolecule e.g. antibiotics, drugs, nutraceuticals, cardiovascular,hepatoprotective, neuro-tonics etc. The present invention has directimplication in drastically reducing the dosage of antibiotics, drugs andanti-cancer agent while increasing the efficiency of absorption ofbioactive molecules.

BACKGROUND OF THE INVENTION

In Ayurveda cows urine is suggested for improving general health. But itis never scientifically tested for any utility alone. The applicantshave developed the curiosity about this component in the preparationsand asked many questions to them; whether the component cow urine ishaving any activity by itself or it does not have any activity butenhance the activity of other components in the preparations? What arethe components present in the urine of cow? Whether the urine containsmicroorganism, which are beneficial? Whether the degradation productsfrom the urine are beneficial? To answer these questions the applicantobtained “Kamadhenu Arka” the urine distillate from “Go-VigyanaAnusandhana Kendra” Nagpur, India. This is the urine distillatesuggested for drinking to improve the general health and sold anddistributed in different size bottles. The applicants tested the urineon Luria agar and broth in sterile condition and did not notice anygrowth of microorganism. To test whether this is inhibitory to growth ofdifferent microorganism, Escherichia coil and Mycobacterium smegmatiswere grown at different temperatures ranging from 20 to 40° C. inpresence and in absence of the cow urine distillate, no significantdifference in the colony count is noticed. Surprisingly, the samedistillate enhanced the antibiotic action on these bacteria leading tothis invention. The novelty of the invention lies in the fact revealedthrough precise experimentation that the enhancement action and itseffectiveness is achievable only in the range of concentration which isliterally in nano to micro molar levels. And when a higherconcentration/dosage is used in the formulation or combinations theactivity(ies) do not appear. That should be the reason for non-detectionsuch a valuable potential of cow urine (Go-mutra). From million doses ofannual antibiotics consumption goes waste as these could not be utilizedor targeted to the infective organisms effectively due to variousfactors like efficient absorption, transportation to the target site,retention time, operation of efflux pump, metabolism etc. Thus, largeportions of the drugs we apply are wasted and only a minisculepercentage is being targeted to the infective microbes. Also, theunutilized drug/antibiotic amount remains as a load in the body andenvironment acting as a selection pressure to facilitate emergence ofdrug resistance in parasites and their predominance, ultimately leadingto failure of antibiotics against resistant infections. This also isresponsible for side effects, illness and reduction in life expectancybeing more acute in the older population. One of the ways, which hasbeen feasible to reduce drug dosage, has been synergism between twotherapeutic agents. However, if both have the antibiotic property, stillthe problem of continued selection pressure on microbes is likely tocontinue. So, the applicants thought of utilizing cow urine, which isnot microbicidal but when present with a drug or active molecule,enhance its activity and availability (bioenhancers). This way, theselection pressure will be counter-balanced simultaneously reducing thedosage of antibiotics or drugs for minimizing the side effects, whichhas also high commercial importance.

The present invention was the result of planned experiments to provide anovel method for improving activity and bioavailability of antibiotics,drugs and other molecules using ‘cow urine distillate’ in differentformulations.

The bioavailability of nutrients and enhancement antibiotics effect isrelevant to human, plant as well as animal health and thus thecompositions and methods of the invention are also intended to be usedin agriculture and veterinary practice.

DESCRIPTION OF RELATED ART

Cow's urine (Go-mutra) can be considered as the most effective animalorigin substance/secretion with the capacity of general healthimprovement but it does need substantiation through scientificexperimentation. Thus, the applicants considered it worthwhile toscientifically look at this and define the molecular basis of the valuesthrough in vitro and in vivo assays. The applicants in the firstinstance probed whether it contained any drug facilitator elements sincesuch a property would make it a highly useful natural substance. Inrecent days, use of ‘piperine’ as a bioavailability enhancer has beendescribed (U.S. Pat. Nos. 5,616,593 and 5,972,382). Till today thus, theknown bioavailability enhancer documented is piperine and a series ofinventions related to this compound have been described in the followingprior arts.

OBJECTS OF THE INVENTION

The main objective is to provide new use of the bio-active fraction as abio-enhancer and as a bioavailability facilitator.

In another objective of the invention is to provide method for improvingactivity and bioavailability of antibiotics, drugs and other moleculesusing active fraction from cow urine distillate.

Still another objective of the invention is to provide a process for theextraction of the active fraction form the cow urine.

SUMMARY OF THE INVENTION

The invention relates to new use of a known abundantly available cowurine distillate as an enhancer of antibiotic action on the target. Themolecule of invention helps in the absorption of antibiotics across thecell membrane in animal cells, gram positive and gram negative bacteriaSimilar activities can also be obtained by using the distillate of theurine of cow at 40–50° C. and from the concentrate, which is lyophilizedand dissolved for further use. Further the urine distillate frombuffalo, camel, deer provides similar activity of bioavailability.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWING

FIG. 1 represents HPLC characters of cow urine (Go-mutra) distillate

DETAILED DESCRIPTION OF THE INVENTION

Our emphasis here was to search a plentifully available material withbioenhancing action of higher potency. Additionally, a property that theapplicants searched was the bioenhancement of a scarcely availableanti-cancer natural agent ‘taxol’ (peclitaxel) which is produced inmicroscopic amounts by the Yew tree (Taxus spps.) and hence is always alimited molecule in availability. Cow urine distillate enhanced thekilling activities of different antibiotics on bacteria. More importantwas the obvious enhancement in the cell division inhibitory activityagainst the breast cancer cell line MCF-7.

In an embodiment of the present invention a pharmaceutical compositioncomprising an effective amount of cow urine distillate as abioavailability facilitator and a pharmaceutically acceptable additivesselected from anticancer compounds, antibiotics, drugs, therapeutic andnutraceutic agents, ions and similar molecules which are targeted to theliving systems.

In another embodiment, the cow urine distillate is used asbioavailability facilitator for anticancer therapy directly or incombination with anticancer molecules.

In still another embodiment, the cow urine distillate is used inantifungal therapy for fungal infections.

In yet another embodiment, the antifungals are azoles, clotrimazole,mystatin, amphotericin and similar materials.

In yet another embodiment, fungi covering infections are mycetial,candida, yeast or other fungicidal compounds.

In still another embodiment, the cow urine distillate is used in TBtherapy including multi drug resistant tuberculosis in combination withisoniazid and other anti-tubercular agents.

In yet another embodiment, the bioavailability facilitator helps intransferring the compound across the membrane and for better effectivityon the target site.

In yet another embodiment, the antibiotics are Quinolones,fluoroquinolones like Nalidixic acid and others like Rifampicin,Tetracycline, ampicillin and similar compounds.

In yet another embodiment, the antibiotics, ions and similar compoundsare isoniazid and hydrogen peroxide.

In yet another embodiment, the bioavailability facilitator helps theantibiotics and other molecules to act better on the target byincreasing the effectivity.

In yet another embodiment, the living system may be bacteria, fungi orany living cells.

In yet another embodiment, the anti bacterial agents are selected fromthe group comprising Quinolones, Rifampicin, Tetracycline andampicillin.

In yet another embodiment, the cow urine (Go-mutra) distillate is in therange between 0.001 μl/ml to 100 μl/ml.

In yet another embodiment, the lyophilized active fraction used is inthe range between 0.1 μg/ml to 100 μg/ml.

In still another embodiment, the bioactive fraction enhances theactivity of anti-bacterial agents, anti-cancer agents andanti-tuberculosis agents from 2 to 80 folds.

In yet another embodiment, the bioactive fraction enhances the activityof anti-bacterial agents from 2 to 80 folds.

In yet another embodiment, the anti bacterial agent is ananti-tuberculosis agent selected from isoniazid, pyrazinamide and othersimilar compounds.

In yet another preferred embodiment, the bioactive fraction enhances theactivity of anti-tuberculosis agents from 2 to 20 folds.

In yet another embodiment of the invention, the anti-cancer agent isselected from group consisting of Paclitaxel (Taxol).

In yet another preferred embodiment of the invention, the bioactivefraction enhances the activity of anti-cancer agents from 2 to 20 folds.

The methodology followed by us for this screening included specificallydesigned bioassays as described below. The bacterial and fungal strainsused in this invention were acquired commercially from Institute ofMicrobial Technology (IMTECH), Chandigarh which possessed correspondingproperties of the ATCC strain mentioned.

1. Assay for Bio-Enhancement of Anti-Infective Agents

-   -   a) The minimum inhibitory concentration (MIC) of antibiotic is        determined against Escherichia coli (equivalent of ATCC 10536),        Bacillus subtilis (equivalent of ATCC 6015) and Mycobacterium        smegmatis (equivalent of ATCC 10231) in broth and disc diffusion        assay.    -   b) The antibiotics agents at concentrations ¼, ⅓, ½ and equal to        MIC are added alone and in combination with the test compound at        varying concentrations on disc and in broth to evaluate the        comparative inhibition.    -   c) These combination showing significant advantage or higher        activity than antibiotic alone in terms of enhanced inhibition        of bacterial growth (large inhibition zone in disc diffusion and        affectivity of lower concentration in broth assay) are picked up        for future testing.    -   d) In broth assay the activity is quantified by counting number        of viable cells in a given treatment and converted in fold        enhancement by combination compared to antibiotic/drug alone in        the killing percentage of cells.    -   e) The pre-treatment assay followed to determine whether the        compound is required along with antibiotic to enhance its        activity or even its withdrawal after treatment or prior to        antibiotic treatment would benefit. For this, the cells are        treated with compound for 4 to 8 hours and then washed free of        it by centrifugation and washing in sterile water. This was        followed by treatment with antibiotic as in steps b to d.        2. Assay for Bio-Enhancement of Anti-Cancer Agents    -   a) MCF-7 (Breast cancer commercial cell line obtained from        National Center for Cell Sciences (NCCS), Pune) is inoculated at        a density of about 0.1×10⁶ cells in MEM medium in the wells of        24 well plate.    -   b) This is replaced with fresh medium after 18 hours in each        well.    -   c) The test component (s) is added at desired concentrations in        different wells just after the medium replacement.    -   d) Observations are recorded on the cell count after 36 hours        for which the following steps are required.        -   i. The medium is removed from the wells.        -   ii. The wells are rinsed with 1 ml PBS (Phosphate buffer            saline).        -   iii. To each well 500 μl of freshly prepared trypsin (0.1%            in PBS) solution is added.        -   iv. Trypsin solution is removed after 30 seconds and the            plate is gently tapped till the cells are released from the            plate surface.        -   v. Fresh 1 ml of MEM growth medium is added and agitated            with a pipette to obtain a cell suspension.        -   vi. 10 μl of cell suspension is taken on the haemocytometer            and a cover glass is placed over the counting chamber.        -   vii. The number of viable cells is counted in 5 big squares            and the readings are taken from 5 microscopic fields to            determine the average.        -   viii. The cell count (titer per ml) in the original sample            is then calculated as average count ×10³.            Composition of Minimum Essential Medium (MEM): 100 ml

MEM powder (Sigma - Aldrich, USA) = 0.96 g HEPES Buffer (Sigma -Aldrich, USA) = 0.26 g Sodium Bicarbonate = 0.22 g Penicillin G =   10mg Streptomycin =   20 mg Gentamycin =   5 mg Foetal Calf Serum =   15ml Foetal Calf Serum =   15 ml Distilled water =   85 ml3. Bioavailability Tests through Biological Membrane

-   -   a) Specially designed U-tubes of glass consisting of two        components (opposite-L type) were used in which one open end of        an L-shaped was tapered to fit within the untapered end of the        other L-tube (FIG. 1).    -   b) The membrane of goat gut (initial part) was stretched and        fixed to act as the barrier between the two ends such that by        joining the two L-tubes, a U-tube was made.    -   c) Sterile distilled water was then filled in both the sides to        equal height/level. The antibiotic/compound was added to the        donor tube (tapered) and through spectro-photometer, the        transfer of molecule was observed using UV and visible        absorption maxima of the respective molecules by taking the OD        at defined wavelengths.

EXAMPLES

In the next step of elucidation of the enhancer action, the applicantsexperimented with the killing activities of different antibioticsagainst the bacteria singly and in combination with the test component(cow urine distillate) following the method described above. Theseexperiments are being described in the following examples. When thebacteria were grown in presence of the compound as such no significantkilling was observed. In all the experiments the cow urine distillateconcentration was kept at 1 μl/mL unless it is specifically mentioned.

Example 1 Cow Urine Distillate Mediated Enhancement in the KillingAction of Antibiotics Against Gram-negative Bacterium Escherichia coli

TABLE 1 Survival fraction of Survival fraction of viable cells uponviable cells upon treatment with *Fold Concentration treatment withantibiotic + cow urine enhancement in Antibiotics μg/ml antibiotic alonedistillate combination antibiotic activity Rifampicin 10 0.86 0.17 5.0Rifampicin 30 0.05 0.007 7.1 Ampicillin 4 1.11 0.60 1.85 Ampicillin 60.09 0.02 4.50 Ampicillin 8 0.05 0.01 5.00

It was calculated as Survival fraction of viable cells upon treatmentwith antibiotic and cow urine distillate in combination/Survivalfraction of viable cells upon treatment with antibiotic alone

Example 2 Cow Urine Distillate Mediated Enhancement in the KillingAction of Antibiotics Against Gram-positive Bacterium Bacillus subtilis

TABLE 2 Survival fraction of Survival fraction of viable *Fold viablecells upon cells upon treatment with enhancement in Concentrationtreatment with antibiotic + cow urine antibiotic Antibiotics μg/mlantibiotic alone distillate combination activity Rifampicin 0.005 1.10.28 5.5 Rifampicin 0.05 0.03 0.01 3.0 Ampicillin 0.1 1.00 0.3 3.3Ampicillin 0.5 0.18 0.06 3.0

Example 3 Cow Urine (Go-mutra) Distillate Mediated Enhancement in theKilling Action of Antibiotics Against Bacterium Mycobacterium smegmatis

TABLE 3 Survival fraction of Survival fraction of viable *Fold viablecells upon cells upon treatment with enhancement in Concentrationtreatment with antibiotic antibiotic + cow urine antibiotic Antibioticsμg/ml alone distillate combination activity Rifampicin 0.05 0.008 0.000180 Rifampicin 0.1 0.006 0.0036 1.5 Ampicillin 0.5 0.07 0.006 11.6

Example 4 Cow Urine Distillate Mediated Enhancement in the KillingAction of Izoniazid and Hydrogen Peroxide Against oxyR Mutant ofBacterium Escherichia coli. The Cow Urine Distillate Concentration is0.001 μl/ml

TABLE 4 Survival fraction of viable cells upon Fold Survival fraction oftreatment with a enhancement viable cells upon isoniazid/H₂O₂ + cow intreatment with urine distillate isoniazid/H₂O₂ Concentrationisoniazid/H₂O₂ alone combination activity Isoniazid 250 μg/ml  7.5 × 10⁷0.99 × 10⁷ 7.5 Hydrogen 0.003% v/v 7.14 × 10⁷  1.2 × 10⁷ 5.9 peroxide(H₂O₂)

The oxyR gene is required for the induction of a regulon of hydrogenperoxide-inducible genes in Escherichia coli (Christman M F, Storz G andAmes B N (1989) Oxy R, a positive regulator of hydrogenperoxide-inducible genes in Escherichia coli and Salmonella typhimurium,is homologous to a family of bacterial regulatory proteins (Proc. Natl.Acad. Sci. (USA). 86:3484–3488.). The mutants of these genes aresensitive to drugs like isoniazid and hydrogen peroxide, which producefree radicals, damaging the cellular systems. So the killing activitiesof these compounds are increased by 5 to 8 folds by cow urine.

Example 5 Cow Urine Distillate Mediated Enhancement in the Activity ofAnti Cancerous Compounds. The Cow Urine Distillate Concentration is 1μl/ml

TABLE 5 Final titre of viable cells upon Taxol Initial titre Final titreof viable treatment with Concentration of viable cells upon treatmenttaxol + cow urine μg/ml cells with taxol alone distillate 0.001 0.9 ×10⁻⁶ 0.059 × 10⁶ 0.039 × 10⁶ 0.005 0.9 × 10⁻⁶ 0.042 × 10⁶ 0.032 × 10⁶0.01 0.9 × 10⁻⁶ 0.036 × 10⁶ 0.012 × 10⁶

The applicants observed similar results of enhancement in the animalcell culture (cancerous cell line MCF-7 obtained from National Centerfor Cell Sciences (NCCS), Pune), in which the killing action of anticancerous chemical ‘Taxol’ is increased. The components of cow urinedistillate in our study help in transferring other compounds across themembrane thereby increasing the absorption in, irrespective of bacteria,animal and plant cell. This in-turn has immense importance forabsorption of the drugs, pharmaceuticals, nutraceutical and otherrelated compounds and ions by the cells.

Example 6 Bioenhancement of Antifungal Agent Clotrimazol by BioactiveFraction Gm-IV. The Concentration of the Active Fraction is Kept at 10μg/ml.

TABLE 6 Minimum inhibitory Concentration (MIC) in Treatment μg/ml Foldenhancement Clotrimazol 4.40 0.0 Clotrimazol + Gm IV 0.88 5.0

In other observations the compound cow urine distillate enhances thetransport of antibiotics e.g. Rifampicin, Tetracycline, Ampicillinacross the gut as well as artificial membrane. The enhancement intransport is approximately 2 to 7 folds.

Great emphasis now is being laid towards quality assurance of crudedrugs from alternative sources widely used in the Indian system ofmedicine. The present invention enlarges the scope and use of the cowurine distillate in therapeutical and nutraceutical application.

Example 7

Development of powder form: For enhancing the utility and convenience ofapplication of cow urine (Go-mutra), the applicants further fractionatedto reach a solid form which is also free of the typical smell of cowurine distillate that it is more readily acceptable to the humans. Forthis purpose the urine distillate was fractionated as described by thefollowing procedure:

Fractionation of a White Crystalline Solid from Cow Urine.

-   Step 1: Cow urine is collected aseptically in the stainless steel    container directly from the cow, which is maintained in hygienic    environment-   Step 2: Fifteen liters of cow urine is distilled continuously at    40–50 ° C. in glass distillation apparatus to obtain 10–12 liters of    the distillate in 16 to 18 hours.-   Step 3: The distillate is packed in surface sterilized plastic or    glass container for further use-   Step 4: 200 ml of cow urine distillate was mixed with half the    volume of methanol and extracted with hexane.-   Step 5: The hexane fraction (Gm-I) was lyophilized and tested for    similar activity as that of cow urine (Go-mutra).-   Step 6: The aqueous fraction was extracted with ethyl acetate and    the ethyl acetate fraction (Gm-II) was lyophilized and tested for    similar activity as that of cow urine (Go-mutra).-   Step 7: Further, aqueous fraction containing white precipitate was    extracted with butanol and the butanol fraction (Gm-III) having pale    yellow precipitate was lyophilized and tested for similar activity    as that of cow urine (Go-mutra).-   Step 8: The remaining aqueous fraction containing white crystalline    precipitate (Gm-IV) was dried and tested for similar activity as    that of cow urine (Go-mutra).

The white powder (Gm-IV) that was obtained in the range of 10 to 20grams per 100 ml of the distillate showed all the above activities asdescribed for cow urine distillate at concentration 0.001 to 10 g/ml,with much more stability and being devoid of the unpleasant smell andhence was used as the advanced product of the invention. The novelty ofthe invention lies in the fact revealed through precise experimentationthat the enhancement action and its effectiveness is achievable only inthe range of concentration which is literally in nano to micro molarlevels. And when a higher concentration/dosage is used in theformulation or combinations the activity(ies) do not appear.

Physical Characters of the Gm-IV Fraction

-   Color: White-   Physical state: Solid Crystalline-   Solubility: Water-soluble and mixture containing water-   Melting point: Above 400° C.-   Specific Gravity: 1.006-   RF value in methanol: Chloroform (50:50) phase: 0.65    HPLC Characterization of Cow Urine (Go-mutra) Distillate

HPLC was performed on LC-8A Shimadzu HPLC with mobile phase water:acetonitrile (80:20), flow rate 10. m/min, UV detection at 275 nm andC-18 E MERCK (150×4 mm) column. Two major peaks (retention time 5.334and 11.310 min) observed in the profile of cow urine (Go-mutra)distillate (FIG. 1).

Further characterization to test the chemical nature of the compound wasperformed through Feigl's test (In: E Stahl, Thin Layer Chromatography)which was positive indicating the presence of glycoside or sugar.

The novelty of the invention is that from cow urine (Go-mutra)distillate, a stable solid fraction could be isolated which is watersoluble and devoid of the urine smell and can directly be used in anyformulation.

The fraction Gm-IV also enhances the transport of antibiotics andvitamins across the mammalian gut membrane. The example describing theenhanced transport of rifampicin by the fraction Gm-IV is given below.

Example 8 Fraction Gm-IV of Cow Urine (Go-mutra) Distillate MediatedEnhancement in the Bioavailability Across the Biological Membrane(Rifampicin, 1 mg/ml and Fraction Gm-IV, 1.0 μg/ml).

TABLE 7 Wave OD measured as Absorbance (specific to the compoundCompound(s) in length maxima) across the membrane in receiving tubeafter the donor tube (nm) 1 hr 2 hr 3 hr 4 hr 5 hr 6 hr Rifampicin A₃₄₀0.0097 0.0214 0.0334 0.0771 0.0858 0.0910 A₄₇₅ 0.0177 0.0284 0.03090.0412 0.0484 0.0496 Rifampicin + Gm-IV A₃₄₀ 0.0525 0.0961 0.1353 0.16390.1919 0.1989 A₄₇₅ 0.0502 0.0904 0.0793 0.0966 0.1157 0.1183

As cow urine (Go-mutra) distillate and the fraction Gm-IV, both showsenhanced membrane permeability by enhancing the antibiotics across thesemi-permeable membrane and mammalian gut membrane, it can be assumedthe above substances can be used for enhancing intestinal transport andtransport of molecules across membranes of various biological functionsincluding urinary/renal systems.

In all the experiments 1 to 5, the enhancing activity of the lyophilizedproduct of the invention is found to have same enhancing activity likethat of the distillate.

The invention can further be explained as follows:

-   -   1. The sample shows enhancement of bioavailability of rifampicin        (antibiotic) and Vitamin B-12 across the mammalian gut membrane        (Goat intestine was used) within 2 hours and it keeps increasing        up to 4 hours.    -   2. It shows clear inhibition of ascorbic acid action to prevent        oxidation of cut apple indicating that it probably enhances the        isoniazid (INH) by oxidative mechanism that synergises the drug        action of INH.    -   3. The distillate showed enhancement action for INH even at        10–50 thousand-fold dilution in the final volume of culture.    -   4. Still more interesting observation is that at 1 μl/ml the        distillate showed enhancement in the activity of taxol by at        least 5 folds. Further experiments in this direction have been        taken up on priority.

1. A process of preparing a white powder from cow urine distillate(GM-IV), said process comprising: a) mixing a volume of cow urinedistillate with a volume of methanol, the volume of the methanol beinghalf the volume of the cow urine distillate; b) extracting with hexaneto produce a hexane fraction (GM-I) and an aqueous fraction; c)extracting the aqueous fraction with ethyl acetate to produce an ethylacetate fraction (GM-II) and an aqueous fraction containing a whiteprecipitate; d) extracting the aqueous fraction containing the whiteprecipitate with butanol to produce a butanol fraction containing a paleyellow precipitate (GM-III) and an aqueous fraction with a whiteprecipitate; and e) drying the aqueous fraction containing the whiteprecipitate from step d) to form the white powder from cow urinedistillate (GM-IV).
 2. The process of claim 1, wherein the white powderfrom cow urine distillate is devoid of a cow urine smell.
 3. The processof claim 1, wherein the white powder the following physicalcharacteristics: a solid crystalline form, water solubility, a meltingpoint above 400° C., a specific gravity of 1.006 and an RF value inmethanol:chloroform (50:50) phase: 0.65.
 4. The process of claim 1,wherein the white power has HPLC properties having two major peaks withretention times of 5.334 min and 11.310 min.